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Position: Discussion forums > Bioscience biotechniques discussion forums > Immunohistochemistry In Situ Hybridization forum
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in situ hybridization: all turn blue!!
Posted by: Mco (IP Hidden, New member, 2)
Date: May 24, 2005 10:33AM
I have a problem with my whole mount in situ hybridization that I cannot understand:
after adding BM purple for revelation, embryos turn blue after one hour only. This happens only for recently synthesized probes. With the old ones, I haven't any problem. Can the probe preparation generate something like that? I wonder if my digoxigenin is contaminated or something?
Re: in situ hybridization: all turn blue!!
Posted by: 5'-ATCG (IP Hidden, Unregistered user, )
Date: May 24, 2005 05:01PM
your probe maybe degraded. How big is it? riboprobe or oligo?
Re: in situ hybridization: all turn blue!!
Posted by: Mco (IP Hidden, New member, 2)
Date: May 25, 2005 12:54AM
My probe is a RNA probe 500bp long. As usual I have checked my probe by running an agarose gel, and it's very ok. I had already synthetized this probe from the same plasmid preparation, and I know it works well.
Re: in situ hybridization: all turn blue!!
Posted by: 5'-ATCG (IP Hidden, Unregistered user, )
Date: May 25, 2005 11:06AM
if there is no signal that may be the DIG issue.
I think either your antibody or blocking, washing condition go wrong....
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